Dynamic allostery in substrate binding by human thymidylate synthase.
NMR
allostery
conformational entropy
cooperativity
human
molecular biophysics
protein dynamics
structural biology
thymidylate synthase
Journal
eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614
Informations de publication
Date de publication:
06 10 2022
06 10 2022
Historique:
received:
02
05
2022
accepted:
31
08
2022
entrez:
6
10
2022
pubmed:
7
10
2022
medline:
12
10
2022
Statut:
epublish
Résumé
Human thymidylate synthase (hTS) is essential for DNA replication and therefore a therapeutic target for cancer. Effective targeting requires knowledge of the mechanism(s) of regulation of this 72 kDa homodimeric enzyme. Here, we investigate the mechanism of binding cooperativity of the nucleotide substrate. We have employed exquisitely sensitive methyl-based CPMG and CEST NMR experiments enabling us to identify residues undergoing bifurcated linear 3-state exchange, including concerted switching between active and inactive conformations in the apo enzyme. The inactive state is populated to only ~1.3%, indicating that conformational selection contributes negligibly to the cooperativity. Instead, methyl rotation axis order parameters, determined by
Identifiants
pubmed: 36200982
doi: 10.7554/eLife.79915
pii: 79915
pmc: PMC9536839
doi:
pii:
Substances chimiques
Nucleotides
0
Thymidylate Synthase
EC 2.1.1.45
Banques de données
Dryad
['10.5061/dryad.wh70rxwqk', '10.5061/dryad.j9kd51cfx']
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM144348
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA016086
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM127698
Pays : United States
Organisme : NIGMS NIH HHS
ID : P41 GM111135
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM083059
Pays : United States
Informations de copyright
© 2022, Bonin et al.
Déclaration de conflit d'intérêts
JB, PS, AL No competing interests declared
Références
J Mol Biol. 2013 Jul 10;425(13):2372-81
pubmed: 23648838
Chem Rev. 2009 Sep;109(9):4108-39
pubmed: 19522502
J Biomol NMR. 2015 Oct;63(2):187-99
pubmed: 26271302
J Am Chem Soc. 2015 Nov 18;137(45):14260-3
pubmed: 26517288
J Biomol NMR. 1995 Nov;6(3):277-93
pubmed: 8520220
J Am Chem Soc. 2004 Mar 31;126(12):3964-73
pubmed: 15038751
Biomol NMR Assign. 2021 Apr;15(1):197-202
pubmed: 33486616
Biochemistry. 2008 Apr 22;47(16):4636-43
pubmed: 18370400
Biochemistry. 1995 Feb 7;34(5):1469-74
pubmed: 7849005
Biochemistry. 1999 Oct 19;38(42):13829-36
pubmed: 10529228
J Am Chem Soc. 2012 May 16;134(19):8148-61
pubmed: 22554188
Biochem Biophys Res Commun. 2013 Jul 12;436(4):725-9
pubmed: 23791741
Bioinformatics. 2005 Jun 15;21(12):2856-60
pubmed: 15827080
Proc Natl Acad Sci U S A. 2017 Jun 20;114(25):6563-6568
pubmed: 28584100
J Biol Chem. 2001 Apr 27;276(17):14170-7
pubmed: 11278511
Angew Chem Int Ed Engl. 2019 May 6;58(19):6250-6254
pubmed: 30847985
J Am Chem Soc. 2013 Oct 9;135(40):15092-100
pubmed: 24007504
Proc Natl Acad Sci U S A. 2007 May 15;104(20):8311-5
pubmed: 17494761
J Biomol NMR. 1994 Sep;4(5):603-14
pubmed: 22911360
Biochemistry. 2010 Mar 23;49(11):2475-82
pubmed: 20151707
Protein Sci. 2011 Jan;20(1):87-94
pubmed: 21064161
Nat Struct Mol Biol. 2006 Sep;13(9):831-8
pubmed: 16906160
J Biol Chem. 2017 Aug 11;292(32):13449-13458
pubmed: 28634233
Proc Natl Acad Sci U S A. 2009 Oct 27;106(43):18249-54
pubmed: 19828436
Biochemistry. 2016 Oct 11;55(40):5702-5713
pubmed: 27649373
Nature. 2018 Jun;558(7709):324-328
pubmed: 29875414
Biosci Rep. 2014 Dec 23;34(6):e00168
pubmed: 25423174
J Am Chem Soc. 2005 Jun 8;127(22):8214-25
pubmed: 15926851
Front Biosci. 2004 Sep 01;9:2521-6
pubmed: 15353304
Nature. 2007 Dec 6;450(7171):913-6
pubmed: 18026087
J Comput Biol. 2013 Oct;20(10):805-16
pubmed: 23992298
J Biomol NMR. 2012 Jul;53(3):209-21
pubmed: 22689066
Biophys J. 2019 Sep 17;117(6):1074-1084
pubmed: 31500803
Proc Natl Acad Sci U S A. 2017 Apr 25;114(17):4424-4429
pubmed: 28348247
J Biomol NMR. 2010 Oct;48(2):65-70
pubmed: 20694505
J Biomol NMR. 2013 Dec;57(4):333-52
pubmed: 24170368
Bioorg Chem. 2010 Jun;38(3):124-31
pubmed: 20199796
J Biomol NMR. 2007 May;38(1):79-88
pubmed: 17464570
J Biol Chem. 2002 Nov 8;277(45):43126-36
pubmed: 12192007
Eur Biophys J. 1984;11(2):103-9
pubmed: 6544679
J Am Chem Soc. 2003 Aug 27;125(34):10420-8
pubmed: 12926967
ACS Chem Biol. 2021 Dec 17;16(12):2766-2775
pubmed: 34784173